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ACS Infect Dis. 2017 Aug 11;3(8):575-584. doi: 10.1021/acsinfecdis.7b00052. Epub 2017 Jun 9.

Optimized Fluorescence Complementation Platform for Visualizing Salmonella Effector Proteins Reveals Distinctly Different Intracellular Niches in Different Cell Types.

Author information

1
Department of Chemistry and Biochemistry, BioFrontiers Institute, UCB 596, University of Colorado , 3415 Colorado Avenue, Boulder, Colorado 80303, United States.

Abstract

The bacterial pathogen Salmonella uses sophisticated type III secretion systems (T3SS) to translocate and deliver bacterial effector proteins into host cells to establish infection. Monitoring these important virulence determinants in the context of live infections is a key step in defining the dynamic interface between the host and pathogen. Here, we provide a modular labeling platform based on fluorescence complementation with split-GFP that permits facile tagging of new Salmonella effector proteins. We demonstrate enhancement of split-GFP complementation signals by manipulating the promoter or by multimerizing the fluorescent tag and visualize three effector proteins, SseF, SseG, and SlrP, that have never before been visualized over time during infection of live cells. Using this platform, we developed a methodology for visualizing effector proteins in primary macrophage cells for the first time and reveal distinct differences in the effector-defined intracellular niche between primary macrophage and commonly used HeLa and RAW cell lines.

KEYWORDS:

Salmonella; bacterial effector proteins; host−pathogen interface; live cell imaging; split-GFP

PMID:
28551989
PMCID:
PMC5720895
DOI:
10.1021/acsinfecdis.7b00052
[Indexed for MEDLINE]
Free PMC Article

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